Thurincin H is a bacteriocin produced by Bacillus thuringiensis with activity against a wide range of bacteria, including Gram positive and Gram negative. Disadvantages of producing thurincin H in B. thuringiensis is the low production level in the native strain probably due to the highly regulated mechanism of biosynthesis. The present study aimed to increase the synthesis of thurincin H produced by the native strain (Btm) through the establishment of additional copies of the structural gene (i.e. thnA) and the genes responsible for the bacterial self-immunity (thnRDE). Here, three recombinant strains of Btm were generated, harboring three, six and nine additional copies of thnA, and three with one copy of thnRDE upstream to the thnA copies. Data showed that the highest yield was obtained at 16 h using three additional copies of thnA (Btm/pHT-One) with a bacteriocin activity of 20,000 U/mg compared with the parental strain which showed 10,000 U/mg, increase of 100% in the production of the bacteriocin. Also, the addition of the genes responsible for self-immunity showed that recombinant B. thuringiensis (Btm/pHT-TwoRDE) can support six additional copies of thnA with an increase of 60% compared with the parental strain.
Keywords: Bacillus thuringiensis; Improving yield; Thurincin H; thnA genes.